Mechanism of Artemisia annua L. in GIOP with kidney-yin deficiency based on network pharmacology / 药学实践杂志

Objective To predict and preliminarily verify the potential targets and related signaling pathways of Artemisia annua L. in treating glucocorticoid-induced osteoporosis (GIOP) with kidney-yin deficiency by network pharmacology and in vitro experiments. Methods The pharmacological targets of Artemisia annua L. were obtained from TCMSP database and were converted to gene names through Uniprot database. The target genes of GIOP with kidney-yin deficiency were obtained from GeneCards database, OMIM database and Drugbank database, and the common target genes were obtained by cross analysis with drug target gene. Protein-protein interaction (PPI) network was constructed by String database, and visualization analysis and core targets screening were performed by Cytoscape 3.9.0. All common targets were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis through Metascape database. Finally, the prediction results were verified by in vitro experiments. Results Ninety-eight targets of Artemisia annua L. to GIOP with kidney-yin deficiency were screened, including 17 core genes. The results of GO and KEGG functional enrichment analysis indicated that Artemisia annua L. treating GIOP with kidney-yin deficiency was related to biological processes such as hormonal response, positive regulation of cell death and extracellular stimulation response, et al, as well as signaling pathways such as PI3K/AKT, AGE/RAGE, MAPK and IL-17 et al. The number of genes enriched in PI3K/AKT signaling pathway was the largest. In vitro experiment results showed that Artemisia annua L. promoted the proliferation of osteoblasts damaged by dexamethasone (DEX), increased alkaline phosphatase activity, activated PI3K/AKT pathway, and promoted the phosphorylation of AKT. Conclusion Artemisia annua L. treating GIOP with kidney-yin deficiency has the characteristics of multi-targets and multi-pathway, which could promote the proliferation and differentiation of osteoblasts through multiple pathways. The PI3K/AKT signaling pathway is an important pathway. Artemisia annua L. treating GIOP with kidney-yin deficiency might be related to its ability to promote the PI3K/AKT signaling pathway and promote the phosphorylation of AKT.

Research overview of anti-osteoporosis effects of traditional animal medicines / 药学实践杂志

Osteoporosis is a systemic bone metabolism disease characterized by low bone mass, bone microstructure destruction, increased bone fragility, and easy fracture，which is more common in the elderly. Animal medicine, as an important part of natural medicines, has the characteristics of wide resources, complex chemical components, and broad pharmacological effects. It has been extensively used in the field of anti-osteoporosis. This article summarizes the pharmacological effects and applications of several major animal medicines for osteoporosis, and discusses the existing problems, aiming to provide a reference for the development of animal drugs against osteoporosis.

Effect and mechanism of Bajitianwan on preventing D-galactose-induced osteoblast bone loss / 药学实践杂志

Objective To explore the effect and mechanism of Bajitianwan on preventing D-galactose (D-gal)-induced osteoblast bone loss. Methods Osteoblasts isolated from 24 h old Wistar rats were injured by D-gal and intervened with Bajitianwan extract. The osteoblastic proliferation and differentiation were determined by MTT and alkaline phosphatase (ALP), respectively. The cell reactive oxygen species (ROS) levels were detected by DCFH-DA fluorescent probes. The expression of cellular oxidation-related protein nuclear factor erythroid 2-related factor 2 (Nrf2), phosphorylated protein kinase B (p-AKT), protein kinase B (AKT), heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase 1 (NQO1) were detected by Western blotting. The intranuclear expression of Nrf2 protein was measured by immunofluorescence. Results Bajitianwan extract had significantly increased the osteoblastic proliferation and differentiation, and significantly reduced the intracellular ROS level. Bajitianwan extract had activated the PI3K/AKT pathway via activating the phosphorylation of AKT in osteoblasts, and promoted NQO1 and HO-1 expression. In addition, Bajitianwan had significantly promoted the expression of Nrf2 in the nucleus of osteoblasts, activating Nrf2 signaling pathway, and further promoted bone formation. Conclusion This study confirmed that Bajitianwan could prevent D-gal injured osteoblastic bone loss for the first time. The mechanism might be related to the regulation of oxidative stress associated PI3K/AKT and Nrf2 signaling pathway.

Microscopic identification for roots from 4 medicinal plants of Actinidia genus / 药学实践杂志

Objective To perform microscopic identification for the roots of Actinidia macrosperma C.F. Liang, Actinidia valvata Dunn, Actinidia arguta (Sieb. & Zucc) Planch. ex Miq., Actinidia chinensis Planch., and provide the basis for judging medicinal materials exactly. Methods The powder microscopic characteristics of 4 medicinal plants of Actinidia genus were observed by microscopic identification method. Results Taking the morphological characteristics of calcium oxalate clusters, starch granules and ducts as the main differences, a key table was compiled to identify the roots of these four medicinal plants. Conclusion The microscopic identification method could effectively distinguish 4 Chinese herbs of Actinidia genus, and which is worth further studying.

Study on chemical constituents of Hippocampus trimaculatus Leach / 药学实践杂志

Objective To study the chemical constituents of Hippocampus trimaculatus Leach. Methods After extracted with ethanol, Hippocampus trimaculatus Leach was isolated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, and reversed-phase C18 column chromatography. The structures of compounds were identified by physical and chemical properties, spectral data and literature comparison. Results Eight compounds were isolated from Hippocampus trimaculatus Leach and identified as L-phenylalanine (1), alanine (2), inosine (3), cholesterol (4), N-acetyltyramine (5), uracil (6), D-mannitol (7), tetrodoine (8), respectively. Conclusion Compounds 5, 7, 8 are isolated from Hippocampus trimaculatus Leach for the first time.

Review of bone metabolism regulated by intestinal flora and the application of Traditional Chinese Medicine / 药学实践杂志

As a complex and large microbial community colonized in the human body, the intestinal flora and its metabolites short-chain fatty acids (SCFAs) play an important role in participating in human metabolism, resisting pathogens, and regulating immune mechanisms. In recent years, many studies have found that the intestinal flora is closely related to bone metabolism. The intestinal flora is able to regulate bone metabolism and affect bone mass changes through various pathways such as absorption of nutrition, generation of SCFAs, regulation of immunity, and influence on body metabolism. The potential pathways and mechanisms by which intestinal flora affect bone mass changes were reviewed in this article in bone metabolism. The related study on Traditional Chinese Medicine that has effects in balancing intestinal flora for regulating bone metabolism was also introduced in order to provide new ideas for the prevention and treatment of osteoporosis, a disease related to bone metabolism.

Effect of traditional Chinese medicine Syngnathus on D-galactose-induced learning and memory impairment in aging mice / 药学实践杂志

Objective To study the effect of traditional Chinese medicine, Syngnathus on learning and memory impairment induced by D-galactose in aging mice and its mechanism of action. Methods HPLC was used to determine the content of DHA, the active ingredient in anti-learning and memory impairment in Syngnathus. The aging mouse model was prepared by intraperitoneal injection of D-galactose (D-gal). Morris water maze test and Western blot were used to detect the ability of learning and memory, biochemical indicators and protein expression related to oxidative damage in the hippocampus, and to explore the protective effect and mechanism of Syngnathus on learning and memory impairment in aging mice. Results HPLC results showed that the DHA content in Syngnathus was 7.761 3 mg/g (calculated as crude drug), accounting for about 47% of the total composition. Morris water maze results showed that Syngnathus could reduce the escape latency of learning and memory-impaired aging mice and increase the target quadrant swimming time, the proportion of swimming distance and the number of times of crossing the platform, and improve the learning and memory impairment of mice. In addition, Syngnathus can activate the AKT/FOXO1/SOD2 signaling pathway in the hippocampus of aging mice with learning and memory impairment, promote the expression of oxidative stress pathway-related proteins, and improve the learning and memory impairment in aging mice by reducing the degree of oxidative damage in the hippocampus of aging mice. Conclusion This study found that Syngnathus is rich in DHA, which has the effect of improving learning and memory impairment induced by D-galactose in aging mice, and preliminarily clarified that its mechanism of action is related to anti-oxidation. Experimental evidence is provided.

Hops extract alleviates Aβ-injury to osteoblasts through antioxidant pathway / 药学实践杂志

Objective To explore the effects of Humulus lupulus L. extract (HLE) and its mechanism on improving bone formation of Aβ-injured osteoblasts. Methods Osteoblasts isolated from 24 h-old Wistar rats were injured by Aβ1-42 oligomer and intervened with HLE. The proliferation, differentiation and bone mineralization of osteoblasts were determined by MTT assay, alkaline phosphatase (ALP) activity assay and alizarin red staining, respectively. The apoptosis of osteoblasts was detected by flow cytometer. The expression levels of bone formation related proteins, and proteins of Nrf2 and FoxO1 pathways were measured by Western blotting analysis. The intranuclear expression of FoxO1 protein was detected by immunofluorescence. Results HLE significantly improved the cell proliferation, ALP activity and bone mineralization, and inhibited the apoptosis of Aβ-injured osteoblasts. HLE also significantly promoted the expressions of collagen type Ι (COL-I) and osteopontin (OPN) in Aβ-injured osteoblasts. HLE notably activated the Nrf2 and FoxO1 signaling pathways in Aβ-injured osteoblasts by promoting the expressions of related proteins and maintained bone metabolism through relieving oxidative stress. Conclusion This study confirms that HLE can alleviate Aβ-injury to osteoblasts, and preliminarily clarifies the mechanism being related to antioxidation, which provides a new reference for the mechanism research and drugs development for anti-osteoporosis.

Effects of Humulus lupulus L. and its active ingredient xanthohumol on preventing glucocorticoid-induced osteoporosis / 药学实践杂志

Objective To explore the effects of Humulus lupulus L. extract (HLE) and xanthohumol (XN) on preventing glucocorticoid-induced osteoporosis (GIOP). Methods The GIOP model was established by intraperitoneal injection of dexamethasone (DEX). Bone microstructure, bone mineral density and serum biochemical indexes were evaluated by Micro-CT and ELISA kits. The levels of cells proliferation and ALP activity, and the expression of bone formation related proteins were assayed with primary osteoblasts injured by DEX. Results HLE and XN significantly alleviated the bone microstructure damage, enhanced the bone mineral density, and improved the trabecular parameters in GIOP mice. In vitro experiments showed that HLE and XN can prevent bone loss not only by improving cell proliferation and ALP activity, but also through increasing the expression of bone γ-glutamic acid-containing proteins (BGP), bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx-2). Conclusion This study confirmed that HLE and XN had anti-GIOP effects for the first time. It provides a new resource for the development of anti-osteoporosis medications.

Application of edible Chinese medicine in epidemic prevention / 药学实践杂志

Edible Chinese medicine is a significant part of traditional Chinese medicine. With the concept of "disease prevention", it can be used in all stages of epidemic prevention and control. This paper introduced the current status of anti-epidemic applications of edible medicines in detoxifying by heat-clearing or blood-cooling, removing dampness and turbidity, Qi and Yin tonifying, etc. In addition, new suggestions and strategies were provided for the professionals in this aera.

Study on the protective effect of menatetrenone against the oxidative stress of osteoblasts / 药学实践杂志

Objective To investigate the protective effect of menatetrenone (MK4) on the osteoblasts in oxidative stress, and to clarify the anti-osteoporosis mechanism of MK4. Methods Mouse osteoblasts (MC3T3-E1) induced by hydrogen peroxide (H2O2) was used. Cell viability, ALP activity and the area of bone nodule were observed. The level of ROS was detected by DCFH-DA, mitochondrial membrane potential by JC-1, apoptosis rate by annexin V-FITC/PI, and the expression of FoxO1, FoxO3, SOD, bcl-2 and bax by RT-PCR. Results Menatetrenone at 10 μmol/L significantly increased the proliferation of osteoblasts stimulated by H2O2, ALP activity, bone nodule formation area, cell membrane potential, the antioxidant SOD and transcription factors FoxO1 and FoxO3 mRNA expression. In the meantime, the elevated malondialdehyde and reactive oxygen species level in cells induced by H2O2, the apoptosis rate and the mRNA expression level of bax/Bcl-2 were significantly reduced. Conclusion menatetrenone can protect osteoblasts from oxidative damage by regulating FoxO pathway and reduce osteoblasts apoptosis by up regulating the proportion of Bcl-2/bax.

Application and prospect of Humulus lupulus L. in anti-osteoporosis / 药学实践杂志

Humulus lupulus is a kind of special resource plant used both as medicine and food in Xinjiang. In addition to being widely used in beer brewing, Humulus lupulus has long been recognized for its medicinal value, especially for the treatment of postmenopausal osteoporosis. Modern pharmacological research shows that its active components have great potential in the development of anti-osteoporosis drugs. However, in recent years, the wild Humulus lupulus resources in China have been seriously degraded, and the contents of active components are quite different. Ensuring high-quality Humulus lupulus germplasm resources is a prerequisite for the development and utilization of Humulus lupulus. This paper reviews the major chemical components of Humulus lupulus and their effects and application in the prevention and treatment of osteoporosis, and discusses the existing problems, aiming to provide a reference for the development and utilization of Humulus lupulus against osteoporosis.

Process optimization of Morinda officinalis with wine steaming by star dot design-response surface methodology / 药学实践杂志

Objective To establish the best wine steaming process for morinda officinalis with monotropein as indicator. Methods Response surface methodology was used to optimize the wine steaming process for morinda officinalis with the amount of rice wine, stewing time, moistening time and the monotropein content as evaluation indexes. Results The best condition was identified with rice wine (rice wine/herbs, g/g) 10%, moistening time 1.0 h, fully steamed and dried. Conclusion The Star dot design-response surface method can be used to optimize the wine steaming process for morinda officinalis.

Effects of vitamin K on osteoblastic bone formation and osteoclastic bone absorption / 药学实践杂志

Objective To compare the effects of vitamin K1 (VK1), vitamin K2 (MK4), vitamin K2 (MK7) and vitamin K3 (VK3) on bone formation and bone absorption. Methods Osteoblasts were isolated from calvaria of newborn rats and osteoclasts were induced by receptor activator of nuclear factor-κ B ligand (RANKL). ALP and TRAP activity were measured by diphenyl phosphate method. Osteoclast metabolic activity was measured by Celltiter kit. The inhibition of cathepsin K (CTSK) was measured by Z-FR-MCA fluorescent substrate and collagen substrate degradation. Results MK4 and MK7 at 0.1~1 μmol/L significantly increased the proliferation of osteoblasts (P<0.05) and at 1 μmol/L increased ALP activity and bone nodule formation area. VK3 inhibited bone nodule formation (P<0.05). VK1,VK3,MK4 and MK7 at 1 μmol/L had no effect on osteoclastic bone absorption. MK4 and MK7 significantly inhibited TRAP activity at 0.1~1 μmol/L (P<0.05), while VK1 and VK3 did not show the inhibitory effect. The inhibition of MK4 at 25 μmol/L on CTSK binding to Z-FR-MCA substrate activity is 58.9% and the inhibition of MK4 at 100 μmol/L on collagen degradation of CTSK activity is 73.2%. Conclusion Compared with VK1 and VK3, MK7 and MK4 significantly increase osteoblast activity and inhibit osteoclast bone absorption, MK4 inhibits osteoclast CTSK enzyme activity.

Research on chemical components and biological activities of the iridoids in Morinda genus / 药学实践杂志

Morinda genus of Rubiaceae has been widely used in medicine at home and abroad. Many parts of Morinda tree are utilized in research, mainly including roots, stems, leaves, branches and seeds. Through the research of online databases, the chemical components and biological activities of the iridoids in Morinda genus were summarized in this paper. Up to now, more than 50 kinds of iridoids have been identified. In addition, more and more studies proved that Morinda iridoids might benefit human via such anti-inflammatory, antinociceptive, anti-oxidation, anti-tumor and bone protection. The theoretical basis was provided for the further development and utilization of the iridoids in Morinda genus.

Establishment of HPLC Fingerprint and Its Spectrum-effect Relationship Study of Humulus lupulus / 中国药房

OBJECTIVE：To establish HPLC fingerprint of Humulus lupulus ，and to investigate its correlation with the antioxidant activity. METHODS ：The determination was performed on Diamonsil C 18 column with mobile phase consisted of 0.1％ phosphoric acid solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 30 ℃，and detection wavelength was set at 358 nm，with sample size of 2 μL. Using xanthohumol as reference，HPLC fingerprints of 11 batches of H. lupulus were determined. The similarity of 11 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition）to confirm common peaks. Using scavenging rate of DPPH and ABTS free radical as pharmacodynamic indicators of antioxidant effects ，SIMCA 14.1 analysis software was used for PLSR to establish the spectra-effect relationship ，and validation test of in vitro anti-oxidation was carried out. RESULTS ：There were 19 common peaks in HPLC fingerprints of 11 batches of sample ，the similarity of which was higher than 0.830. Seven components were identified as xanthohumol，cohumulone，humulone，adhumulone，colupulone，lupulus and adlupulus. Eleven batches of H. lupulus had the ability to scavenge DPPH and ABTS free radicals. The spectrum-effect relationship showed that xanthohumol ，humulone and colupulone peaks were positively associated with its anti-oxidant ability ，and variable projection value was greater than 1. In vitro antioxidant results showed that scavenging effect of 0.1 mg/mL xanthohumol to DPPH free radicals was similar to that of 0.01 mg/mL vitamine C ，but scavenging effect of 10.0 mg/mL xanthohumol to ABTS free radical was less than that of 0.1 mg/mL vitamin C. CONCLUSIONS：Established HPLC fingerprint can be used for the quality evaluation of H. lupulus ；xanthohumol，humulone and colupulone are the main material basis for the antioxidant effect of H. lupulus

Study on fingerprint of Hupulus lupulus by HPLC / 药学实践杂志

Objective To establish fingerprint of Hupulus lupulus by HPLC.Methods Diamonsil C18column (250mm×4.6mm, 5μm) was used.The mobile phase consisted of acetonitrile and 0.1％phosphoric acid, in gradient elution mode, detection wavelength was 358nm, column pressure at 10.2MPa, column temperature was 30℃, injection volume of 10μl and flow rate was 1.0ml/min.Results A common mode of HPLC fingerprint of Hupulus lupulus was established.19common peaks were calibrated, xanthohumol, cohumulone, humulone, adhumulone, colupulone, lupulone and adlupulone for identification.Among the 26batches of Hupulus lupulus, 5batches of herbs had similarities of less than or equal 0.9, which were all wild varieties.Conclusion The method was fast, effective, accurate and reliably to distinguish the cultivation and wildness Hupulus lupulus, which provided reference for quality control of Hupulus lupulus.

Anti-osteoporotic study on xanthohumol, an active ingredient in Humulus lupulus L / 药学实践杂志

Objective To evaluate the anti-osteoporotic effect of xanthohumol(XN)in animal and osteoblast.Methods The anti-osteoporotic study on XN was performed with ovariectomized mice model.Serum biochemical indexes,bone mineral density(BMD)and bone histomorphology were measured using Elisa kits and Micro-CT analysis.In vitro test,the effect of XN on osteoblastic proliferation,differentiation and mineralization were assayed.The expression of protein related to bone for-mation was measured by Western blot analysis.Results In vivo experiments,XN significantly increased the estrogen level, reduced the high bone turnover rate,improved the microenvironment and BMD in ovariectomized mice.In vitro experiments, XN protected bone loss not only by promoting osteoblastic proliferation,ALP activity and bone mineralization,but also through increasing the expression of osteopontin(OPN),bone sialoprotein(BSP)and bone morphogenetic protein-2(BMP-2). Conclusion This is the first report to confirm that XN has anti-osteoporotic effect,which provides a new approach for the clin-ical treatment of osteoporosis.

Analysis of flavonoids contents of hops / 药学实践杂志

Objective To determine the contents of total flavonoids and xanthohumol in hop from 29 different countries and regions .Methods Rutin colorimetric method was used to determine the content of total flavonoids .HPLC-UV method was established for the determination of xanthohumol in hops .HPLC method was performed by Dikma Technologies Diamonsil C18 (250 mm × 4 .6 mm ,5 μm ) column with mobile phase acetonitrile-1％ glacial acetic acid solution at the flow rate of 1 .0 ml/min .The column temperature was 25 ℃ .The detection wavelength was 370 nm .Results The equation of linear re-gression of total flavonoids was A=30 .345C+0 .0168 ,r=0 .9999 .The equation of linear regression of xanthohumol was A=55446 C+9040 .5 ,r=0 .9999 .Their linear ranges were respectively 20 .2-404 .0 μg/ml ,2 .152-43 .040 μg/ml ,which indica-ted a good linear relationship .The RSDs of precision and repeatability were less than 2％ .The average recoveries of flavonoids and xanthohumol were respectively 102 .71％ and 100 .21％ .Conclusion The contents of total flavonoids and xanthohumol in different hops varieties are significantly different and the import hops was better than the domestic hops in this study .

Study on the technology for extracting lignans in schisand ra chinensis (Turcz ) Baill by AB-8 macroporous resin / 药学实践杂志

Objective To study the conditions for isolating and purifying lignans in schisandra chinensis (Turcz) Baill by AB-8 type of macroporous adsorption resin .Methods The content of Schizandrol ,Deoxyschizandrin ,r-Schisandrin was deter-mined by HPLC .The optimum separation and purification process of lignans from schisandra chinensis (Turcz) Baill with AB-8 macroporous adsorption resin were identified by static and dynamic adsorption ,desorption tests .The sample volume ,concen-tration ,eluting velocity ,and ethanol concentration were investigated .Results The optimum conditions for isolating and purif-ying lignans in schisandra chinensis (Turcz) Baill by AB-8 macroporous adsorption resin were as follows :dosage of sample liq-uid was 1BV ,concentration of sample solution was 9 .159-16 .523 mg/ml ,ratio of diameter to height of resin column 1:5 ,ad-soption flow rate was 2 .5 BV/h ,eluted by 5 BV 30％ ethanol ,followed by 10BV 95％ ethanol .The transfer rate of lignans was 77 .07％ with 22 .06％ of total lignan content .Conclusion AB-8 macroporous adsorptive resin can effectively isolate and purify lignans from schisandra chinensis (Turcz) Baill .This low cost process is easy to operate and with high industrial pro-duction value .